tbuckley/cabot-search huggingface dataset:
Abstract P2004: T Lymphocytes Promote Diastolic Dysfunction And Fibrosis In A Non-Hypertensive Angiotensin II Mouse Model Of Heart Failure
Authors: Michelle M Kim, Eleonora Mezzaroma, Nadia Martinez Naya, Adolfo G Mauro, Pilar Alcaide, Antonio Abbate, Stefano A Toldo
Journal: Circulation Research (2023)

Background: Angiotensin (Ang) II signaling and diastolic dysfunction are hallmarks of heart failure (HF). Ang II promotes hypertension (HTN)-mediated cardiac fibrosis and remodeling as well as T lymphocyte activation during the pathogenesis of HF. We hypothesized that AngII induces HF, independent of HTN, through T cell activation in a mouse model. Methods: Using a non-HTN AngII model, we administered AngII (0.2 μg/kg/day) or H 2 O vehicle through subcutaneous pumps in wild type (WT) or T cell receptor α knockout ( Tcrα -/- ; bereft of T cells) mice (15 wks old, N=5-7/group). We used flow cytometry to detect T cells (CD45+CD3+) in LV single-cell suspensions. LV dimensions, ejection fraction (EF), and diastolic function (isovolumetric relaxation time - IRT) were measured after 4 wks with Doppler echocardiography. LV end-diastolic pressure (EDP) was measured with a Millar catheter. Heart slides were stained with picrosirius red to measure interstitial fibrosis. Results: T cells were significantly increased in the LV by AngII vs H 2 O (23.0±2.1% vs 9.4±1.7%, p&lt;0.001). All groups had similar LV dimensions and EF (not shown). AngII increased IRT (15.1±1.0 vs 8.6±0.6 ms, p&lt;0.0001) and LVEDP (9.0±1.3 vs 3.2±0.5 mmHg, p&lt;0.001) in WT mice vs H 2 O, but did not increase IRT and LVEDP in Tcrα-/- mice (11.8±0.8 vs 9.5±0.8 ms and 3.2±0.8 vs 2.9±0.5 mmHg; all p=ns), which were significantly lower than that of WT mice (all p&lt;0.05). AngII increased interstitial fibrosis in WT mice (2.2±0.2 vs 0.7±0.1%, p&lt;0.0001) while Tcrα-/- hearts had significantly less fibrosis (1.0±0.1%, p&lt;0.001). Conclusions: T cells are increased in the LV and promote diastolic dysfunction and fibrosis in a non-HTN AngII HF mouse model.
Abstract P1030: Alpha Kinase 3 Signaling At The M-band Regulates Sarcomeric Protein Quality Networks
Authors: James W McNamara, Benjamin Parker, Holly Voges, Neda Mehdiabadi, Francesca Bolk, Feroz Ahmad, Antonia Zech, Kevin Watt, Enzo R Porrello, David Ellliott
Journal: Circulation Research (2023)

Objectives: Striated muscle contraction is driven by molecular machinery composed of specialized cytoskeletal proteins, called the sarcomere. Pathogenic variants in genes that encode these sarcomeric proteins trigger a heart disease called cardiomyopathy, which causes abnormal growth and function of the heart muscle and can cause sudden death. However, there is a growing appreciation that genetic mutations in genes not encoding the traditional sarcomeric proteins also substantially contribute to cardiomyopathy. One such gene is ALPK3 , encoding an atypical alpha kinase. Despite its strong link to cardiomyopathy, there have been no studies to define its molecular function. Therefore, the objective of our study was to understand the genetic basis of ALPK3- induced cardiomyopathy and define its molecular function in health and disease. Methods &amp; Results: We created a suite of gene-edited human pluripotent stem cells (hPSCs) carrying endogenous fusion proteins, loss of function mutations, or disease-causing patient variants, and differentiated these hPSC lines into human ventricular muscle cells. Using reporter hPSCs, we found that ALPK3 is localized to the M-band of the cardiac sarcomere, in contrast to its posited nuclear localization. ALPK3 deficiency impaired muscle cell function and sarcomere organization in both hPSC cardiac organoids and mice harboring a pathogenic ALPK3 variant. We performed multiomic analysis on ALPK3-deficient hPSC cardiomyocytes to define the function of ALPK3 and observed consistent enrichment of sarcomere and protein quality control pathways. Interactomics demonstrated that ALPK3 links the cytoskeleton to protein surveillance networks at the M-Band. Notably, the ubiquitin-binding protein SQSTM1 bound ALPK3, could be phosphorylated by ALPK3, and was mislocalized from the sarcomere in three independent ALPK3 patient lines. Conclusions: Together, our data demonstrate that ALPK3 regulates the link between the sarcomere and protein surveillance system and that its deficiency impairs cardiac function via this mechanism. This work highlights a potential new pathway to target for therapy development and treatment of patients with cardiomyopathy.
Abstract P3061: Development Of Molecular Targeted Therapy Against Right Ventricular Failure: Involvement In Complement C3- Factor D- C3a Pathway
Authors: Shogo Ito, Shinsuke Yuasa, Hiroyuki YAMAKAWA, Jin Komuro, Hisayuki Hashimoto, Mai Kimura, Eichi Nakao, Keiichi Fukuda, Yoshihiro Fukumoto
Journal: Circulation Research (2023)

Backgrounds: Right ventricular (RV) failure plays a critical role in right heart failure and left heart failure. However, there is no specific therapy developed for RV failure. To elucidate a novel therapeutic target against RV failure, we focus on RV specific pathway. Methods: Microarray analysis using several parts of adult murine heart was conducted and differentially expressed genes (DEGs) were applied to pathway analysis. Molecular mechanism was examined by using neonatal rat ventricular cardiomyocyte (NRVM) in vitro. To understand the function of target molecule in vivo, we induced RV failure by pulmonary artery constriction (PAC) in mice and inhibition experiments were performed using RV failure model. Results: In microarray analysis elucidated 995 genes as DEGs in RV. Pathway analysis showed that alternative complement pathway-related genes were significantly up-regulated in RV. Moreover, complement factor D (Cfd) and C3a was a potential upstream factor attributable to unique feature of RV. Administration of C3a recombinant protein to primary cultured ventricular myocyte indicated that phosphorylation of several MAP kinases were enhanced. In vivo, in C3KO PAC mice, RV dysfunction was significantly suppressed, and histological study showed that RV fibrosis was significantly suppressed. In wild type mice with PAC, administration of C3a receptor antagonist dramatically improved RV dysfunction and reduced RV fibrosis. Additionally, in vivo electrophysiological study revealed that the inducibility of ventricular arrhythmia was increased in wild type PAC mice, but ventricular arrhythmia was significantly attenuated in C3KO PAC mice. Furthermore, electrophysiological analysis by isolated cardiomyocyte showed that calcium overload was also attenuated in RV cardiomyocyte of C3KO PAC mice. Conclusion: We revealed that complement C3a was highly produced in RV and chemical or genetical blockade of C3a ameliorates RV dysfunction and RV fibrosis in PAC mice. C3a was to be a potent bioactive protein for immunocompetent cells that played an important role in modulating cardiac function. Accordingly, we demonstrated that the blockade of C3a had a potential role for novel therapeutic target to RV failure.
Abstract P1144: Single-cell RNA-Sequencing Of The Murine Cardiac Conduction System Reveals Transcriptional Regulators Associated With Conduction Development And Disease
Authors: Benjamin Beyersdorf, Felix Atzert, Sean M Wu, William R Goodyer
Journal: Circulation Research (2023)

Rhythmic beating of the heart is driven by a small subset of heart cells, altogether known as the cardiac conduction system (CCS). The CCS consists of unique components including the sinoatrial node (SAN), the atrioventricular node (AVN), the bundle of His, bundle branches and Purkinje fiber (PF) system. Due to obstacles inherent in studying the CCS, including small cell numbers, large cell-type heterogeneity and complex anatomy, our understanding of the transcriptional networks driving CCS development remains limited. Here, we harnessed single-cell RNA-sequencing (scRNAseq) and bioinformatic analyses to uncover putative DNA-binding transcription factors (TFs) as well as transcriptional co-regulators enriched within the entire developing murine CCS. Comparison of multiple independent prior datasets, spanning all major components of the CCS, revealed previously known TFs necessary for conduction development including, Islet 1 ( Isl1 ; avg log fold change 0.34, p= 1E-192) in the SAN; T-box transcription factor 5 (Tbx5 ; avg log fold change 0.54, p= 2.1E-110) in the AVN; and ETS variant transcription factor 1 ( Etv1; avg log fold change 0.89, p= 6.7E-105) in the PFs, among others. Notably, an additional 16 novel DNA-binding transcription factors and 40 transcription co-regulators were enriched within specific CCS components at statistical significance. To further evaluate the transcriptional landscape of individual subcomponents within the SAN (compact versus transitional cells), we also performed scRNAseq on FACS-isolated SAN cells from embryonic day 16.5 Hcn4 -GFP transgenic mice revealing differential expression of a host of TFs, both established and novel. Finally, our murine findings were then cross-referenced to over 15 human GWAS studies related to cardiac rhythm abnormalities, revealing an association of a subset of these murine CCS-enriched transcription factors and cardiac arrhythmias in humans. In conclusion, this study provides a comprehensive gene expression atlas of transcription factors and transcriptional co-regulators within individual CCS components at single-cell resolution, laying the groundwork for future studies into the transcriptional regulation of cardiac conduction development and disease.
Abstract P3189: Vcam-1-mediated Mitochondrial Biogenesis In Macrophages Exacerbates Inflammation And Atherosclerosis
Authors: Niranjana Natarajan, Jonathan Florentin, Ebin Johny, Lee Ohayon, Partha Dutta
Journal: Circulation Research (2023)

Atherosclerotic cardiovascular disease is a leading cause of adult mortality worldwide. Macrophages play a key role in atherogenesis by sustaining the local inflammatory response, facilitating accumulation of immune cells in atheromas that contribute to plaque development. Mitochondrial metabolism plays a key role in governing inflammatory gene expression in macrophages. Yet, the role of mitochondrial biogenesis and mitochondrial DNA synthesis in regulating macrophage inflammation is not known. Vascular cell adhesion protein 1 (VCAM-1) expressed by endothelial cells mediates monocyte adhesion and extravasation in developing atherosclerotic plaques. We observed increased VCAM-1 expression in human and murine plaque macrophages. However, the function of this cell adhesion molecule expressed by macrophages is not known. Here, we evaluated if and how myeloid-VCAM-1 drive mitochondrial biogenesis, inflammation, and atherosclerosis pathology. Increased VCAM-1 expression in plaque macrophages correlated with oxidative DNA damage and mitochondrial volume. To understand the role of myeloid-Vcam1 in atherogenesis, we generated Apoe -/- LyzM cre/+ VCAM-1 fl/fl mice. Consistently, Apoe -/- mice lacking Vcam1 in macrophages exhibited reduced atherosclerosis severity and inflammation. Vcam1-deficient macrophages exhibited decreased inflammation, oxidative phosphorylation, mitochondrial biogenesis, and mitochondrial DNA synthesis genes, including Cmpk2. Cmpk2 deletion in macrophages after oxidized LDL treatment reduced inflammatory mediators that aggravate atherosclerosis. RNA sequencing analysis of Vcam-1 -deficient plaque macrophages and analysis of macrophages lacking Cmpk2 identified Fcor and Lyz1 as the target genes of Vcam1 and Cmpk2 . Interestingly, atherosclerotic plaque macrophages deficient of Sting , which mediates inflammatory signaling in response to oxidized mitochondrial DNA, had increased levels of Fcor and Lyz1 . Our data suggest that VCAM-1 in macrophages signals via CMPK2 and POLG to promote mitochondrial biogenesis, oxidation, and fragmentation, mediating atherogenesis.
Abstract P3097: Sex-related Differences In Immune Responsiveness Of Sarcospan-deficient Mice To Metabolic Stress
Authors: Luaye Samarah, Isela C Valera, Gisienne Reis, Aida Rahimi Kahmini, Bolade Olateju, Nazanin Mohammadipour, Michelle S Parvatiyar
Journal: Circulation Research (2023)

This study compares sex-related differences in inflammatory signaling that may contribute to metabolic and cardiovascular disease. The SSPN gene encoding the sarcospan (SSPN) protein has been described as an “obesity-susceptibility” gene. To study the influence of SSPN on metabolic function we subjected young 2-month-old male and female WT and sarcospan-deficient (SSPN -/- ) mice to 4 months high fat diet (HFD) (60% fat). Young male and female SSPN -/- mice exhibited some protection from weight gain and tissue inflammation. Cardiac function was preserved in the SSPN -/- mice of both sexes after HFD. Thus, we became interested in the combined impact of SSPN and metabolic stress on systemic inflammation. To study how metabolic stress impacts activation of anti- and pro-inflammatory pathways the chemokine/cytokine/adipokine proteome was examined in blood serum, heart, white adipose tissue, and liver of male mice. Under control conditions both WT and SSPN -/- male blood serum exhibited low levels of pro-inflammatory molecules, however CXCL 10 and CCL-2 significantly increased in SSPN -/- after HFD. Histological analysis of white adipose tissue (WAT) further revealed that SSPN deficiency may be protective against WAT immune infiltration in both males and females. Proteome analysis of WAT from HFD and control diet males revealed striking alterations in inflammatory signatures, whereas in the heart and liver most significant changes were observed in SSPN -/- controls. Flow cytometry was performed to examine spleen immune cell content of the mice. To elucidate factors contributing to the sex differences observed in the SSPN -/- response to metabolic stress we examined the response of bone marrow derived macrophages (BMDM) obtained from the sex/diet/gender groups included in this study. The BMDM acute response to innate immune ligands was tested at 3 and 6 hours to innate immune ligands and in general, female control diet SSPN -/- BMDMs exhibited enhanced pro-inflammatory responsiveness compared to female HFD and male control diet BMDMs. Overall, these studies describe distinct inflammatory signatures in the heart and WAT and a sex-dependent influence of SSPN -/- on macrophage activation that may have distinct consequences on cardiovascular and metabolic health.
Abstract P3051: Master Splicing Regulator Rbpms2 Is Not Required For Murine Cardiac Development Or Contractile Function
Authors: Jared M McLendon, Xiaoming Zhang, Ryan L Boudreau
Journal: Circulation Research (2023)

Post-transcriptional gene regulation (PTGR) through RNA-binding proteins (RBP) is an understudied area of cardiac biology. Emerging research describes important roles for Rbpms and Rbpms2 in RNA splicing, cardiomyocyte diploidy and proliferation, cytoskeletal reorganization and translation initiation during cardiomyocyte differentiation and cardiac development. However, the role of Rbpms2 in adult heart is largely unknown. We identified Rbpms2 as a cardiovascular-enriched RBP that is consistently downregulated in failing hearts from humans and mice. Though prior research has focused on roles in splicing, a large portion Rbpms2 protein is extranuclear, and &gt;35% of empirically determined Rbpms binding sites localize in 3’UTRs, suggesting alternative PTGR roles. To assess Rbpms2’s role in cardiomyocyte PTGR, we assessed mRNA-seq changes in neonatal rat cardiomyocytes after overexpression and knockdown of Rbpms2. Pathway analyses revealed significant upregulation of genes involved in cell cycle control, and downregulation of genes involved in viral host defense pathway after Rbpms2 knockdown. We next characterized Rbpms2 knockout (KO) mice, which are viable without overt phenotypes, to provide initial descriptions of adult mouse cardiac function after Rbpms2 deletion. Baseline echocardiography data of male mice at 6 months of age shows normal cardiac size, structure, and function without significant difference between KO and wildtype (WT) mice (n=10). To assess the role of Rbpms2 in cardiac hypertrophy, we induced pressure overload via transverse aortic constriction (TAC) on mix-sex WT and Rbpms2-KO mice, which resulted in robust hypertrophic responses and reduced ejection fraction in both groups, but no significant differences between WT and KO mice (n=11 female, n=7-10 male). Overall, our findings are surprising given recently published data showing that Rbpms2 is a master regulator of RNA splicing affecting sarcomere organization and cardiomyocyte contractile function in zebrafish and stem cell-derived cardiomyocytes, supporting the need for follow-up studies on cardiac Rbpms2. It is possible that Rbpms compensates for the loss of Rbpms2 in the mouse heart and would require double KOs to assess synergistic phenotypes.
Abstract P3077: Withdrawal Of High Fat Diet And L-NAME Dampens T Cell Activation And Ameliorates Pathology In Experimental Heart Failure With Preserved Ejection Fraction (HFPEF) Through Modulation Of The T Cell IRE1a-XBP1 Axis
Authors: Sasha Smolgovsky, Abraham Bayer, Kuljeet kaur, Mark Aronovitz, Erin Sanders, Robert M Blanton, Edward B Thorp, Gabriele G Schiattarella, Joseph A Hill, Juan R Cubillos-Ruiz, Pilar Alcaide
Journal: Circulation Research (2023)

Introduction: Cardiometabolic HFpEF is characterized by T cell systemic inflammation and can be modeled preclinically with high fat diet (HFD) and L-NAME. Metabolic perturbations can alter T cell function through modulation of the unfolded protein response (UPR). How T cells are activated in HFpEF and whether T cell inflammation can be tuned by removing cardiometabolic stress remains unknown. Hypothesis: HFD and L-NAME induce reversible alterations in T cell UPR signaling that modulate T cell dependent cardiac inflammation and pathology in experimental HFpEF. Methods: Male C57/BL6J mice (wild-type, WT), OT-II antigen restricted mice, and CD4 Cre Xbp1 fl/fl mice were fed a high-fat diet (HFD) and L-NAME (H/L) for 5 weeks. WT mice were switched to standard diet (STD) for two additional weeks. WT mice receiving HFD or L-NAME only, or TAC surgery (a model of HF with reduced EF) were used as controls. Left ventricular (LV) end diastolic pressure volume relationship (EDPVR) was assessed using invasive hemodynamic analyses and LV EF by echocardiography. Cardiac and splenic T cells were characterized by flow cytometry and T cell UPR was evaluated by qPCR and Western blot. Results: 5 weeks of H/L was sufficient to induce cardiac T cell infiltration and splenic T cell activation. OTII mice fed H/L showed increased cardiac CD4 + T cell infiltration compared to STD controls, indicating T cell activation is not cardiac antigen restricted. Furthermore, splenic T cell characterization revealed H/L resulted in decreased phosphorylation of the UPR protein inositol-requiring enzyme 1α (IRE1α) and reduced expression of X box-binding protein 1 ( Xbp1s/u ) compared to T cells from mice exposed to HFD or L-NAME alone, STD, or WT TAC controls. CD4 Cre+ Xbp1 fl/fl mice (T cell Xbp1 -/- ) fed with H/L had increased cardiac T cell infiltration compared to Cre - controls. Lastly, Xbp1s expression in splenic CD4 + T cells was recovered after withdrawal of H/L and resulted in partial reduction of cardiac hypertrophy, decreased EDPVR, as well as less cardiac T cell infiltration compared to mice kept on H/L. Conclusions: Reversible downregulation of the T cell IRE1α-XBP1 axis is a unique T cell signature of HFpEF that contributes to T cell inflammation and adverse cardiac remodeling.
Abstract P1057: Outcomes Of Impella Versus ECMO In Multivessel Percutaneous Coronary Intervention
Authors: Yaser Khalid, Kirti Dasu, Hiral Amin, Nielsen Gabriel, Sanath Shetty, Neethi Dasu, Debesh Jha, Tyler Klecha
Journal: Circulation Research (2023)

Background: Impella is a mechanical circulatory support device indicated in severe cardiogenic shock and high-risk percutaneous coronary intervention. ECMO is also used during the percutaneous coronary intervention (PCI) in an acute coronary syndrome that is complicated by refractory cardiogenic shock. There have been few studies examining short-term mechanical circulatory support devices for patients that undergo PCI, so the choice between Impella and ECMO is still a matter of debate. We investigated the in-hospital outcomes of Impella and ECMO in patients who underwent multi-vessel PCI (MVPCI). Methods: In a retrospective cohort study of National Inpatient Sample 2017 data, we used ICD-10 codes to identify patients who underwent MVPCI and received Impella or ECMO support. Multivariate logistic regression was used to adjust for patient demographics, hospital demographics, and relevant comorbidities. Primary outcomes (inpatient mortality, length of stay (LOS), and total hospital charges. Secondary outcomes, such as respiratory failure (RF), gastrointestinal hemorrhage, cerebrovascular accident (CVA)/transient ischemic attack (TIA), intracerebral hemorrhage (ICH), acute kidney injury (AKI), ventricular tachycardia (VT), and atrial fibrillation, were also analyzed. Results: Among 64,489 patients who had MVPCI, 102 (%) received ECMO support, and 927 (%) received Impella support. Both Impella and ECMO use was associated with high in-hospital mortality (Impella 21.8% vs ECMO (49.5%), high hospital LOS (ECMO 19.9 days vs Impella 9.28 days), and total hospital charges (Impella $3,200,000 vs ECMO $7,500,00). Mortality (OR 4.27, p&lt;0.0001), hospital LOS (OR 5.9, p&lt;0.0001), and total hospital charges (OR 7.52, p&lt;0.0001) for patients undergoing MVPCI. There was no statistically significant difference in complications such as RF, gastrointestinal hemorrhage, CVA/TIA, ICH, AKI, VT, and atrial fibrillation. Hispanic patients had higher mortality with ECMO (OR 2.28, p &lt;0.024) than all other ethnicities and races. Conclusions: Both patients with Impella support and ECMO support had significant inpatient mortality. Our analysis suggests that Impella patients had better outcomes than patients on ECMO for MVPCI.
Abstract P1051: Minimally Invasive Delivery Of Synthetic Cardiac Stromal Cells In A Large Animal Model Of Myocardial Infarction
Authors: Yuan Li, Ke Huang, Ke Cheng
Journal: Circulation Research (2023)

Cell therapies for cardiac tissue regeneration are commonly used for tissue remolding post-MI, however, due to viability, retention, and cost, acellular therapeutic components are being explored. Synthetic cardiac stromal cells (synCSCs) generated by encapsulating secreted factors from isolated human cardiac stromal cells were shown to promote cardiac repair after MI. Previously, synCSCs implantation required surgy. Reducing the risks of surgical complications is essential for translational research. It is beneficial to develop a minimally invasive approach such as intrapericardial cavity (iPC) injections. This pre-clinical study aims to elucidate the therapeutic effects of synCSC embedded in hyaluronic acid (HA) hydrogel on a large animal model of MI through iPC injection. HA hydrogel and synCSCs are fabricated according to previous studies. Video-assisted thoracoscopic surgery is performed for iPC injections in Yorkshire male pigs. Magnetic resonance imaging (MRI) and immunohistochemistry (IHC) data were collected for cardiac function analyses. This pre-clinical study aims to bridge the gap between the bench and bedside by using a minimally invasive approach of iPC injection of synCSCs embedded in the hydrogel. Figure 1 shows iPC injection of synCSCs in HA improved cardiac function and cardiomyocyte proliferation in the pig MI model. Single-cell RNA sequencing data is being analyzed to identify gene markers elevated by synCSC treatment and further uncover the molecular pathways activated by synCSC. iPC injection of synCSC provides a unique dataset in the field of cardiac tissue regeneration and offers another therapeutic option for patients suffering from MI.